CN101555462B - Bacteria agent for deeply processing phenylamine oil refinery effluent and preparation method thereof - Google Patents
Bacteria agent for deeply processing phenylamine oil refinery effluent and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a bacteria agent for deeply processing phenylamine oil refinery effluent by a biological method and a preparation method thereof; the preparation method comprises the followingsteps: totaled 17 grafting aerobic microorganism strains and anaerobic microorganism strains are respectively grafted into 17 prepared triangular flasks, each of which contains 50ml to 150ml of liqui d seed culture medium, according to a proportion of 2 percent to 7 percent by volume; the 17 triangular flasks are all placed in a shaking culture bed with constant temperature ranging from 28 DEG C to 35 DEG C to conduct shaking culture for 4 days to 7 days; the 17 strains in the liquid culture media are respectively grafted to 17 solid extension culture media and then all put in a greenhouse with temperature of 30 DEG C to 35 DEG C for 7 days to 11 days; next, 13 aerobic microorganism solid expansion culture products are mixed to prepare an aerobic microorganism solid bacteria agent, and 4 anaerobic microorganism solid expansion culture products are mixed to prepare an anaerobic microorganism solid bacteria agent. The invention adopts a biological deep processing technology and has low production energy consumption, high recycling rate and high degradation rate, thus being capable of realizing the industrial cleaning and continuous deeply processing of the phenylamine-contained oil refinery effluent.
Description
Technical field
The present invention relates to a kind of bacteria agent for deeply processing phenylamine oil refinery effluent and preparation method thereof.
Background technology
Refinery water belongs to high concentration hard-degraded organic waste water, and bigger to microorganism toxicity, biodegradability is poor, and oil composition complexity in the waste water, and saltiness is big, and potential of hydrogen changes greatly, so intractability is very big.Wherein contained aniline and phenyl amines pollutent advanced treatment be difficulty comparatively, and the contradiction that can't reach new emission standard requirement becomes increasingly conspicuous.Domestic most of oil refining industries all adopt physics, chemical process to be handled at present, but it is higher that these methods are handled energy consumption, the strong acid-bases that adopt in the processing more, residual organic pollutant difficult degradation in the processed waste water, difficult, the difficult utilization easily caused the repeated contamination of water body, is difficult in actual applications promote.
Bioremediation is to handle the most effectual way of phenylamine oil refinery effluent.It is to utilize aerobic and the anaerobion alternating action, makes aniline and amino benzenes compounds obtain deep decomposition.Owing to adopt bioremediation lower than physics, chemical process cost, production energy consumption is low, non-secondary pollution, and microorganism again has very strong severe environment adaptability and very high cyclic utilization rate again, so bioremediation has become the Perfected process of advanced treatment phenylamine oil refinery effluent.
Highly active amino benzenes compounds degradation bacteria is achieved the degree of depth degraded of aniline and amino benzenes compounds.Aniline and the amino benzenes compounds in the degradation property microbiological treatment trade effluent used in present many chemical plant, and obtain unusual effect, but compound complicated component in the contaminate environment, the pH fluctuation is big, and saltiness is big, probably suppresses the growth of degradation bacteria or induces its variation, cause degradation bacteria to environmental pollutants degradation speed slow down, be difficult to reach the requirement of actual needs, contaminated environment can not effectively be kept the biological activity of degradation bacteria, makes degradation bacteria bacterium number be difficult to keep higher level.
Summary of the invention
The object of the present invention is to provide a kind of biological process advanced treatment that adopts to contain microbial inoculum of phenylamine oil refinery effluent and preparation method thereof.
The preparation method of bacteria agent for deeply processing phenylamine oil refinery effluent of the present invention comprises the following steps:
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be in the corresponding respectively triangular flask that inserts 17 50-150ml liquid seed culture mediums that prepare of the ratio of 2%-7%, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 28-35 ℃ constant-temperature shaking culture bed jointly, shaking culture 4-7 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases, aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 30-35 ℃ of greenhouse jointly, cultivated 7-11 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
Aerobic microbiological is belonged to by Corynebacterium, gas pseudomonas bacillus genus, pseudomonas, genus arthrobacter, bacillus and microbot and forming, and the blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-7: 1-7: 1-7: 1-10, and surplus is that solid enlarges culture medium;
Anaerobion is made up of enterobacter, Pseudomonas, bacillus and Microbacterium, and the blending ratio of 4 anerobe solid enlarged culturing things is:
Enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=1-10: 1-10: 1-10: 1-10, surplus is a solid enlarged culturing base, all buys at Research for Industrial Microbial Germ preservation center, Gansu Province.
The composition of liquid seed culture medium and weight percentage are: peptone 0.05-0.3%, yeast extract paste 0.1-0.3%, NaCl0.1-0.5%, sucrose 0.2-0.5%, extractum carnis 0.1-0.3%, KH
2PO
40.01-0.04%, MgSO
40.01-0.03%, CaCl
20.01-0.03% and urea 0.01-0.05%, water is settled to 50-150ml, and the pH value is 6.4-7.0;
The composition and the weight percentage of solid enlarged culturing base are: grain 70-75%, wheat bran 15-22%, Semen Maydis powder 2-3%, peptone 0.05-0.3%, yeast extract paste 0.1-0.3%, NaCl0.1-0.5%, sucrose 0.2-0.5%, extractum carnis 0.1-0.3%, KH
2PO
40.01-0.04%, MgSO
40.01-0.03%, CaCl
20.01-0.03%, urea 0.01-0.05% and water 4.95-9.95%, the pH value is 6.4-7.2.
Just can make product bacteria agent for deeply processing phenylamine oil refinery effluent of the present invention with the inventive method, finished product bacteria agent for deeply processing phenylamine oil refinery effluent of the present invention does not need preservation.
The using method of microbial inoculum: with weight proportion is that the aerobic of 10%-15% is filled to respectively in aerobic bacteria aeration cylinder and the anerobe aeration cylinder with the anaerobism solid fungicide, open aerobic aeration and anaerobism waste water circulation apparatus for continous treatment then, speed Continuous Flow by 0.5ml/min (maximum processing capability of device) adds the refinery water that contains 0-160mg/L aniline and aniline compound, this waste water is flowed through respectively in order and is precipitated tube, the nutrition deployment tube, potential of hydrogen is adjusted tube, the temperature equilibrium tube, elementary aeration cylinder, the aerobic bacteria aeration cylinder, the aerobic bacteria aeration cylinder, potential of hydrogen is adjusted tube, the anerobe circulating barrel, the anerobe circulating barrel, potential of hydrogen is adjusted tube, the aerobic bacteria aeration cylinder, the aerobic bacteria aeration cylinder, totally 13 process cartridges get final product the refinery water that continuous advanced treatment contains aniline category matter.
The invention solves present refinery water handle in the aniline processing efficiency not high and handle after still contain problem such as minor amounts of aniline class difficult degradation pollutent, have following advantage:
1. energy-conserving and environment-protective, energy-conserving and environment-protective, existing phenylamine oil refinery effluent treatment technology adopts the physical chemistry technology more, and its investment is huge,
A large amount of uses of water, electricity, coal and strong acid and strong base simultaneously can cause secondary pollution, destroy ecotope.And the present invention adopts biological process advanced treatment technology, and its production energy consumption is low, and reusable edible rate height can not cause repeated contamination to environment, has long-range ecological benefits.
2. microbial inoculum of the present invention is to come by guiding, domestication, screening that many concentration phenyl amines, nitrobenzene, compound fragrant hydrocarbon and microorganism must nutritive elements, and it is the highest that its biological activity all can reach in containing amino benzenes compounds broad concentration range.
3. microbial inoculum of the present invention is made up of multiple aerobic and anaerobion Pseudomonas, and it has the adaptability of broad on pH, temperature, environment, and the while, the preparation method was simple, easy handling.
4. the degradation rate height adopts microbial inoculum of the present invention, it is implemented in by homemade aerobic aeration and anaerobism circulation apparatus for continous treatment contains in the phenylamine oil refinery effluent, and its degradation rate can reach 95%-100%.
5. easy handling is handled in batch production, adopts the microbial inoculum of patent of the present invention, need only with self-control aerobic aeration and anaerobism circulation apparatus for continous treatment in proportion, the principle amplification, can realize that batch production cleans continuous advanced treatment and contain phenylamine oil refinery effluent.
Embodiment
The following examples can make those skilled in the art more fully understand the present invention, but do not limit the present invention in any way.
When using product bacteria agent for deeply processing phenylamine oil refinery effluent of the present invention to handle waste water, with homemade aerobic aeration and anaerobism circulation apparatus for continous treatment, this device is by the precipitation tube, the nutrition deployment tube, potential of hydrogen is adjusted tube, the temperature equilibrium tube, elementary aeration cylinder, the aerobic bacteria aeration cylinder, the aerobic bacteria aeration cylinder, potential of hydrogen is adjusted tube, the anerobe circulating barrel, the anerobe circulating barrel, potential of hydrogen is adjusted tube, aerobic bacteria aeration cylinder and aerobic bacteria aeration cylinder are formed, volume is 9.4L, wherein the anerobe circulating barrel adopts the built-in dividing plate circulation of totally enclosed, all the other tubes all adopt the stainless steel reel, this device is handled when containing phenylamine oil refinery effluent, and maximum processing capability is 0.5ml/mim.
Below specific implementation method by preparation 1Kg aerobic microbial agent, preparation 1Kg anaerobion microbial inoculum the present invention is described, finished product aerobic microbiological solid fungicide before the mixing that the present invention prepares and the water content of this 17 strain bacterium of anaerobion solid fungicide are respectively 9%-14%.
Embodiment 1
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be that corresponding respectively to insert 17 volumes that the 150ml liquid seed culture medium is housed that prepare be in the triangular flask of 500ml for 2% ratio, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 35 ℃ constant-temperature shaking culture bed jointly, shaking culture 6 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases; Aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 30 ℃ of greenhouses jointly, cultivated 7 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
The blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=8: 8: 8: 7: 9: 8: 8: 8: 8: 7: 7: 7: 10, and surplus is that solid enlarges culture medium;
The blending ratio of 4 anerobe solid enlarged culturing things is: enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=9: 9: 10: 10, and surplus is a solid enlarged culturing base.
Each amounts of components of liquid seed culture medium is: peptone 0.45g, yeast extract paste 0.45g, NaCl0.75g, sucrose 0.75g, extractum carnis 0.45g, KH
2PO
40.06g, MgSO
40.045g, CaCl
20.045g, urea 0.075g, water is settled to 150ml, pH7.0.
Solid enlarged culturing base by each amounts of components is: grain 750g, wheat bran 100g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 99.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 60mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.With weight proportion is that 15% aerobic and anaerobism solid fungicide is filled to respectively in aerobic bacteria aeration cylinder and the anerobe aeration cylinder, open aerobic aeration and anaerobism waste water circulation apparatus for continous treatment then, speed Continuous Flow by 0.5ml/min (maximum processing capability of device) adds the refinery water that contains 60mg/L aniline and aniline compound, this waste water is flowed through respectively in order and is precipitated tube, the nutrition deployment tube, potential of hydrogen is adjusted tube, the temperature equilibrium tube, elementary aeration cylinder, the aerobic bacteria aeration cylinder, the aerobic bacteria aeration cylinder, potential of hydrogen is adjusted tube, the anerobe circulating barrel, the anerobe circulating barrel, potential of hydrogen is adjusted tube, the aerobic bacteria aeration cylinder, the aerobic bacteria aeration cylinder, totally 13 process cartridges, take out mouth of a river water sample, record the aniline density loss to 0.0mg/L, degradation rate can reach 100%.
Comparative Examples 1
Do not dose any microbiobacterial agent in aerobic aeration and anaerobism circulation apparatus for continous treatment, after device is opened, take out mouth of a river water sample, aniline and aniline compound are that 60mg/L drops to 57.3mg/L by starting point concentration, and degradation rate only reaches 4.5%.
Embodiment 2
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be that corresponding respectively to insert 17 volumes that the 100ml liquid seed culture medium is housed that prepare be in the triangular flask of 250ml for 5% ratio, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 33 ℃ constant-temperature shaking culture bed jointly, shaking culture 7 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases; Aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 35 ℃ of greenhouses jointly, cultivated 11 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
The blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=10: 10: 10: 10: 10: 10: 10: 10: 10: 7: 7: 7: 10, and surplus is that solid enlarges culture medium;
The blending ratio of 4 anerobe solid enlarged culturing things is: enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=10: 10: 10: 10, and surplus is a solid enlarged culturing base.
The consumption of liquid seed culture medium is: peptone 0.3g, yeast extract paste 0.3g, NaCl0.5g, sucrose 0.5g, extractum carnis 0.3g, KH
2PO
40.04g, MgSO
40.03g, CaCl
20.03g, urea 0.05g, water be settled to 100ml, pH7.0.
Solid enlarged culturing base is formed by weight and is counted: grain 750g, wheat bran 150g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 49.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 100mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.With weight proportion is that 20% aerobic and anaerobism solid fungicide is filled to respectively in aerobic bacteria aeration cylinder and the anerobe aeration cylinder, open aerobic aeration and anaerobism waste water circulation apparatus for continous treatment then, speed Continuous Flow by 0.5ml/min (maximum processing capability of home-made contrivance) adds the refinery water that contains 100mg/L aniline and aniline compound, take out mouth of a river water sample, record the aniline density loss to 1.4mg/L, degradation rate can reach 98.6%.
Comparative Examples 2
Do not dose any microbiobacterial agent in aerobic aeration and anaerobism circulation apparatus for continous treatment, after device is opened, take out mouth of a river water sample, aniline and aniline compound are that 100mg/L drops to 96.7mg/L by starting point concentration, and degradation rate reaches 3.3%.
Embodiment 3
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be that corresponding respectively to insert 17 volumes that the 50ml liquid seed culture medium is housed that prepare be in the triangular flask of 150ml for 5% ratio, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 33 ℃ constant-temperature shaking culture bed jointly, shaking culture 7 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases; Aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 35 ℃ of greenhouses jointly, cultivated 9 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
The blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1, and surplus is that solid enlarges culture medium;
The blending ratio of 4 anerobe solid enlarged culturing things is: enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=1: 1: 1: 1 surplus is a solid enlarged culturing base.
The consumption of liquid seed culture medium is: peptone 0.15g, yeast extract paste 0.15g, NaCl0.25g, sucrose 0.25g, extractum carnis 0.15g, KH
2PO
40.02g, MgSO
40.015g, CaCl
20.015g, urea 0.025g, water is settled to 50ml, pH7.0.
Solid enlarged culturing base is formed by weight and is counted: grain 750g, wheat bran 150g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 49.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 160mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.With weight proportion is that 20% aerobic and anaerobism solid fungicide is filled to respectively in aerobic bacteria aeration cylinder and the anerobe aeration cylinder, open aerobic aeration and anaerobism waste water circulation apparatus for continous treatment then, speed Continuous Flow by 0.5ml/min (maximum processing capability of home-made contrivance) adds the refinery water that contains 160mg/L aniline and aniline compound, take out mouth of a river water sample, record the aniline density loss to 6.7mg/L, degradation rate can reach 95.8%.
Comparative Examples 3
Do not dose any microbiobacterial agent in aerobic aeration and anaerobism circulation apparatus for continous treatment, after device is opened, take out mouth of a river water sample, aniline and aniline compound are that 160mg/L drops to 156.5mg/L by starting point concentration, and degradation rate reaches 2.2%.
Embodiment 4 microbial inoculums only are made up of aerobic microbiological,
Is that the volume that 5% ratio inserts the 50ml liquid seed culture medium for preparing respectively is in the triangular flask of 150ml with aerobic microbiological in percentage composition by volume, the liquid seed culture medium triangular flask seals with the air-permeable envelope of 2 layer of 10 μ m, place 33 ℃ constant-temperature shaking culture bed, shaking culture 7 days, then with 13 bacterial classification liquid seeds respectively correspondence connect and 13 solid enlarged culturing bases, aerobic solid medium adopts 3 layer of 200 order gauze to cover, place 35 ℃ of greenhouses, cultivated 9 days, then 13 aerobic bacteria solid enlarged culturing things are mixed, promptly make the aerobic bacteria solid fungicide.
The blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1: 1, and surplus is that solid enlarges culture medium.
The consumption of liquid seed culture medium is: peptone 0.15g, yeast extract paste 0.15g, NaCl0.25g, sucrose 0.25g, extractum carnis 0.15g, KH
2PO
40.02g, MgSO
40.015g, CaCl
20.015g, urea 0.025g, water be settled to 50ml, pH7.0.
The consumption of solid enlarged culturing base is: grain 750g, wheat bran 150g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 49.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 160mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.With weight proportion is that 20% aerobic solid fungicide is filled to the aerobic tube in homemade aerobic aeration and the anaerobism circulation apparatus for continous treatment, do not load microbial inoculum in the anaerobism tube, opening device then, speed Continuous Flow by 0.5ml/min (maximum processing capability of home-made contrivance) adds the refinery water that contains 160mg/L aniline and aniline compound, take out mouth of a river water sample, record the aniline density loss to 53.9mg/L, only illustrate and also can handle waste water with the aerobic bacteria solid fungicide, degradation rate reaches 66.3%, and effect is handled waste water not as using aerobic microbiological solid fungicide and anaerobion solid fungicide simultaneously.
Comparative Examples 4
Do not dose any microbiobacterial agent in aerobic aeration and anaerobism circulation apparatus for continous treatment, after device is opened, take out mouth of a river water sample, aniline and aniline compound are that 160mg/L drops to 156.5mg/L by starting point concentration, and degradation rate reaches 2.2%.
Embodiment 5 microbial inoculums only are made up of anaerobion,
Is in the triangular flask of 150ml with anaerobion by 5% volume that inserts the 50ml liquid seed culture medium for preparing respectively, the liquid seed culture medium triangular flask seals film with threeply and seals, place 33 ℃ constant-temperature shaking culture bed, shaking culture 7 days, then with 4 bacterial classification liquid seeds respectively correspondence connect and 4 solid enlarged culturing bases, the anaerobism solid medium adopts 2 layers of kraft paper to cover, place 35 ℃ of greenhouses, cultivated 9 days, then 4 anerobe solid enlarged culturing things are mixed, promptly make the anerobe solid fungicide.
The blending ratio of 4 anerobe solid enlarged culturing things is: enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=1: 1: 1: 1, and surplus solid enlarged culturing base.
The consumption of liquid seed culture medium: peptone 0.15g, yeast extract paste 0.15g, NaCl0.25g, sucrose 0.25g, extractum carnis 0.15g, KH
2PO
40.02g, MgSO
40.015g, CaCl
20.015g, urea 0.025g, water be settled to 50ml, pH7.0.
The consumption of solid enlarged culturing base is: grain 750g, wheat bran 150g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 49.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 160mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.
With weight proportion is that 20% anaerobism solid fungicide is filled to the anaerobism tube in aerobic aeration and the anaerobism circulation apparatus for continous treatment, do not dose microbial inoculum in the aerobic aeration tube, opening device then, speed Continuous Flow by 0.5ml/min (maximum processing capability of home-made contrivance) adds the refinery water that contains 160mg/L aniline and aniline compound, take out mouth of a river water sample, record the aniline density loss to 125.9mg/L, only illustrate and also can handle waste water that effect is handled waste water not as using aerobic microbiological solid fungicide and anaerobion solid fungicide simultaneously with the anaerobion solid fungicide.Degradation rate only reaches 21.3%.
Comparative Examples 5
Do not dose any microbiobacterial agent in aerobic aeration and anaerobism circulation apparatus for continous treatment, after device is opened, take out mouth of a river water sample, aniline and aniline compound are that 160mg/L drops to 156.5mg/L by starting point concentration, and degradation rate reaches 2.2%.
Embodiment 6
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be that corresponding respectively to insert 17 volumes that the 150ml liquid seed culture medium is housed that prepare be in the triangular flask of 500ml for 2% ratio, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 35 ℃ constant-temperature shaking culture bed jointly, shaking culture 6 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases; Aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 30 ℃ of greenhouses jointly, cultivated 7 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
The blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 (GSICC 30337): Corynebacterium HY4 (GSICC 30336): Corynebacterium HY5 (GSICC 30338): the gas pseudomonas bacillus belongs to HY2 (GSICC 30136): the gas pseudomonas bacillus belongs to HY3 (GSICC30137): pseudomonas HY6 (GSICC31603): Arthrobacter B-111 (GSICC30144): bacillus HY9 (GSICC32824): bacillus HY10 (GSICC 32825): phenol 8 (GSICC 32838): phenol 13 (GSICC 32821): phenol 82 (GSICC 32811): microbot belongs to HY7 (GSICC 31301)=8: 8: 8: 7: 9: 8: 8: 8: 8: 7: 7: 7: 10, and surplus is that solid enlarges culture medium;
The blending ratio of 4 anerobe solid enlarged culturing things is: enterobacter YY2 (GSICC 30509): Pseudomonas YY4 (GSICC 31616): bacillus YY5 (GSICC 32812): Microbacterium YY7 (GSICC 31302)=9: 9: 10: 10, and surplus is a solid enlarged culturing base.
The consumption of liquid seed culture medium is: peptone 0.45g, yeast extract paste 0.45g, NaCl0.75g, sucrose 0.75g, extractum carnis 0.45g, KH
2PO
40.06g, MgSO
40.045g, CaCl
20.045g, urea 0.075g, water be settled to 150ml, pH7.0.
The consumption of solid enlarged culturing base is: grain 750g, wheat bran 100g, Semen Maydis powder 30g, peptone 3g, yeast extract paste 3g, NaCl5g, sucrose 5g, extractum carnis 3g, KH
2PO
40.4g, MgSO
40.3g, CaCl
20.3g, urea 0.5g, water 99.5g, pH7.2.
The using method of microbial inoculum: the main component that contains phenylamine oil refinery effluent of processing is: aniline and amino benzenes compounds 160mg/L, peptone 3g/L, yeast extract paste 3g/L, NaCl5g/L, sucrose 5g/L, extractum carnis 3g/L, KH
2PO
40.4g/L, MgSO
40.3g/L, CaCl
20.3g/L, urea 0.5g/L, pH7.2.
With aerobic solid fungicide is that 15% to be inoculated in the volume that 500ml contains aniline and amino benzenes compounds concentration 160mg/L refinery water be in the triangular flask of 1000ml by weight ratio, placed 35 ℃ of 140r/min shaking culture 4 days, and then be 15% to be seeded to wherein by weight ratio with the anaerobism solid fungicide, 35 ℃ leave standstill cultivation 4 days, aniline and amino benzenes compounds concentration are reduced to 11.36mg/L, and degradation rate reaches 92.9%.
Comparative Examples 6
Add 15% distilled water and be used as microbial inoculum in containing aniline and amino benzenes compounds concentration 160mg/L refinery water, aniline and aniline compound are that 160mg/L drops to 158.1mg/L by starting point concentration, and degradation rate reaches 1.2%.
Test-results shows:
1. do not add any microorganism, treatment effect is extremely low, and adds microbiobacterial agent of the present invention, and treatment effect obviously improves;
2. though use the aerobic or anaerobion microbial inoculum among the present invention that certain degradation effect is arranged separately, can not reach the purpose of advanced treatment;
3. be used aerobic and anaerobion microbial inoculum among the present invention and can reach the purpose of advanced treatment phenylamine oil refinery effluent;
4. microbiobacterial agent of the present invention cooperates homemade aerobic aeration and anaerobism circulation apparatus for continous treatment can reach the optimum handling effect.
Claims (2)
1. the preparation method of a bacteria agent for deeply processing phenylamine oil refinery effluent is characterized in that it is made up of the following step:
With aerobic microbiological and anaerobion totally 17 strain bacterium by volume percentage composition be in the corresponding respectively triangular flask that inserts 17 50-150ml liquid seed culture mediums that prepare of the ratio of 2%-7%, the liquid seed culture medium triangular flask of aerobic bacteria seals with the air-permeable envelope of 2 layer of 10 μ m, the liquid seed culture medium triangular flask of anerobe seals film with threeply and seals, place 28-35 ℃ constant-temperature shaking culture bed jointly, shaking culture 4-7 days, then with 17 bacterial classification liquid seeds respectively correspondence connect and 17 solid enlarged culturing bases, aerobic bacteria solid enlarged culturing base adopts 3 layer of 200 order gauze to cover, anerobe solid enlarged culturing base adopts 2 layers of kraft paper to cover, place 30-35 ℃ of greenhouse jointly, cultivated 7-11 days, then with 13 mixed aerobic microbiological solid fungicides that get of aerobic bacteria solid enlarged culturing thing, with 4 mixed anaerobion solid fungicides that get of anerobe solid enlarged culturing thing;
Aerobic microbiological is belonged to by Corynebacterium, gas pseudomonas bacillus genus, pseudomonas, genus arthrobacter, bacillus and microbot and forming, and the blending ratio of 13 aerobic bacteria solid enlarged culturing things is:
Corynebacterium HY1 GSICC 30337: Corynebacterium HY4 GSICC 30336: Corynebacterium HY5GSICC 30338: the gas pseudomonas bacillus belongs to HY2 GSICC 30136: the gas pseudomonas bacillus belongs to HY3 GSICC30137: pseudomonas HY6 GSICC31603: Arthrobacter B-111 GSICC30144: bacillus HY9 GSICC32824: bacillus HY10 GSICC 32825: phenol 8 GSICC 32838: phenol 13 GSICC 32821: phenol 82 GSICC 32811: microbot belongs to HY7 GSICC 31301=1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-10: 1-7: 1-7: 1-7: 1-10, and surplus is that solid enlarges culture medium;
Anaerobion is made up of enterobacter, Pseudomonas, bacillus and Microbacterium, and the blending ratio of 4 anerobe solid enlarged culturing things is:
Enterobacter YY2 GSICC 30509: Pseudomonas YY4 GSICC 31616: bacillus YY5 GSICC 32812: Microbacterium YY7 GSICC 31302=1-10: 1-10: 1-10: 1-10, surplus is a solid enlarged culturing base;
The composition of liquid seed culture medium and weight percentage are: peptone 0.05-0.3%, yeast extract paste 0.1-0.3%, NaCl0.1-0.5%, sucrose 0.2-0.5%, extractum carnis 0.1-0.3%, KH
2PO
40.01-0.04%, MgSO
40.01-0.03%, CaCl
20.01-0.03% and urea 0.01-0.05%, water is settled to 50-150ml, and the pH value is 6.4-7.0;
The composition and the weight percentage of solid enlarged culturing base are: grain 70-75%, wheat bran 15-22%, Semen Maydis powder 2-3%, peptone 0.05-0.3%, yeast extract paste 0.1-0.3%, NaCl0.1-0.5%, sucrose 0.2-0.5%, extractum carnis 0.1-0.3%, KH
2PO
40.01-0.04%, MgSO
40.01-0.03%, CaCl
20.01-0.03%, urea 0.01-0.05% and water 4.95-9.95%, the pH value is 6.4-7.2.
2. the bacteria agent for deeply processing phenylamine oil refinery effluent that makes according to the described preparation method of claim 1.
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